Bioinformatics: The collection, processing, and analysis of biological information and data using

computer software.
Gene Technology: A procedure by which one or more selected genes are removed from one
organism and inserted into another, which is then said to be transgenic.
Gene Therapy: Treatment of a genetic disorder by altering a persons genotype.
Genetic profiling (fingerprinting): Sequencing a length of DNA from an organism, to compare with
the sequence of the same DNA from other organisms.
Genetic screening: Testing an embryo, fetus, or adult to find whether a particular allele is present.
Recombinant DNA (rDNA): DNA made by joining pieces from two or more different sources.
Sticky ends: short lengths of unpaired bases. They can easily for hydrogen bonds with
complementary sequences of bases on other pieces of DNA cut with the same restriction enzyme.
Vector: The means of delivering genes to a cell, used in genetic technology.
Plasmid: Small, circular pieces of double-stranded DNA which occur naturally in bacteria and often
contain genes for antibiotic resistance.
Promoter: A length of DNA to that controls the expression of a gene by allowing RNA polymerase
to bind to it as it starts transcription.
Allozymes: Variant forms of enzymes which are produced by different alleles of the same gene.
Variable number tandem repeat (VNTR): A length of DNA that contains different numbers of
repeated base sequences in different individuals in a species.
DNA probe: A single stranded DNA with a sequence complementary to the desired sequence.

Gene transfer basics

- Genetic engineering allows us to overcome barriers to gene transfer between species. Unlike in
selective breeding, where whole sets of genes are involved, with gene technology one single
gene can be transferred on its own. These are the steps of gene transfer:
1. The gene is identified. It is either cut from a chromosomes, made from mRNA by reverse
transcriptase, or synthesised from nucleotides.
2. Polymerase chain reaction (PCR) is used to make many copies of the gene.
3. The gene is inserted into a vector (e.g. plasmid) which delivers the gene into cells.
4. The vector and thus the gene are taken up by the cell.
5. The cells with the new gene are identified and cloned.

It is now possible to synthesise nucleotides artificially, since we have sequenced many proteins.
Scientists simply choose the cons which code for the specific amino acid sequences they want. A
computer holds all this information and can direct the synthesis of short fragments of DNA, which
can then be inserted into plasmids for use in genetic engineering.This method can be used instead
of reverse transcription and cutting out chromosomal DNA. It is used to generate new genes, which
can be used in the synthesis of vaccine or in making the genomes of bacteria.
Tools necessary:
- Enzymes: Restriction endonuclease, ligase, reverse transcriptase
Restriction enzymes: They are a class of enzymes from bacteria which recognise and break
down the DNA of invading viruses known as bacteriophages.
- They are called restriction enzymes because they restrict viral infections.
- They bind to specific restriction sites (a specific sequence of bases) on DNA and make
cuts there. The restriction sites are usually palindromic (can be read the same way
backwards and forwards.
- They do not target bacterial DNA because that DNA is protected either by chemical
markers or not having those restriction sites.
- Restriction enzymes either cut to give blunt ends (cut straight across the sugar-
phosphate backbone) or sticky ends (staggered, diagonal cut).
- When long pieces of DNA are cut with a restriction enzyme, many different lengths of
DNA are formed. To find a certain length of DNA, electrophoresis is necessary.
DNA ligase: used to link together the sugar-phosphate backbones of the DNA molecule and
the plasmid.
- Vectors: Plasmids, viruses, and liposomes
Using plasmids for genetic engineering:
1. The bacteria containing the plasmid are treated with enzymes to break down their cell
walls.
2. The naked bacteria are then spun at high speed in a centrifuge, so that the relatively
large bacterial chromosomes are separated from the small plasmids.
3. The circular DNA of the plasmid is cut using a restriction enzyme.
4. The same restriction enzyme is used to cut the gene, so that the sticky ends are
complementary. If blunt ends are formed, sticky ends must be added.
5. DNA ligase links together the DNA molecule and the plasmid, producing a closed circle
of double-stranded DNA containing the new gene.
6. The bacteria are treated by being placed in a solution of a high concentration of calcium
ions, are cooled and then are given a heat shock to increase the chances of the plasmids
passing through the cell surface membrane.
- A small proportion (about 1%) of the bacteria take up the plasmids with the gene (are
transformed). The rest either don't take up the plasmid or take up a plasmid that closed
without the gene.
Bacterial plasmids can be modified to produce good vectors. They can also be made
artificially (e.g. pUC).
- pUC have a low molecular mass, so they are readily taken up by bacteria.
- They have an origin of replication so they can be copies.
- They have several single target sites for different restriction enzymes in a short length
of DNA (polylinker).
- They have one or more marker genes which allow cells, which have taken them up to
be identified.
- Genes coding for easily identifiable substances which can be used as markers. These genes are
added to the plasmid and are identifiable because:
They lead to resistance to antibiotics.
- The insulin gene can be introduced into the plasmid at the point in the gene for tetracycline
resistance. Any bacteria taking up this plasmid with recombinant DNA will not be able to
survive in agar containing tetracycline.
- This method has fallen out of favour because of fears that antibiotic resistance may spread
to other bacteria. Although this is unlikely due to the isolated environment of the GM
bacteria, the method has decreased in popularity.
They make a fluorescent protein which is easily seen.
- A gene is transferred into the plasmid from a jellyfish.
- It produces a green fluorescent protein (GFP). It fluoresces under ultraviolet light.

They produce an enzyme whose action we can identify.

- Beta-glucuronidase (GUS), found in E. coli, can transform specific colourless compounds
or non-fluorescent substrates into coloured or fluorescent products.
- The enzyme lactase converts a colourless substrate into a blue product.